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Blood, Vol. 109, Issue 7, 2982-2984, April 1, 2007
CpG oligodeoxynucleotides allow for effective adoptive T-cell therapy in chronic retroviral infection
Blood Kraft et al.
109: 2982
Supplemental materials for: Kraft et al, Vol 109, Issue 7, 2982-2984
Files in this Data Supplement:
- Table S1. IFNγ production by virus-specific CD8+ T cells after cocultivation with CpG-stimulated FV-infected DCs (PDF, 51 KB) -
To obtain FV-infected DCs, bone marrow–derived DCs were enriched for expression of the glycosylated FV gag protein using flow cytometry. DCs (5 × 103) were incubated for 72 hours with 5 × 104 naive TCRtg CD8+ T cells. DCs were stimulated with 6 µg/mL ODNs prior to coculture. All assays were performed in duplicate with similar results. IFN concentrations in cell-culture supernatants were measured by enzyme-linked immunosorbent assay (BD OptEIA; BD Bioscience, Heidelberg, Germany).
- Figure S1. CpG-ODN treatment at different time points after adoptive transfer (PDF, 276 KB) -
Chronically FV-infected (B10 × A.BY) F1 mice were treated with 4 × 106 TCR-transgenic, virus-specific CD8+ T cells. B-type CpG-ODN 1668 (CpG; 15 nmol) or control ODNs without CpG motif (Control) were injected at different time points after cell transfer. At 1 week after transfer, the spleen cells of the recipients were harvested, and host (CD45.1–; bottom right) and donor (CD45.1+; top right) CD8+ T cells were stained for IFN, granzyme B (gzmB), and CD107a. Representative flow data are shown for each group. Harvested cells were restimulated for 5 hours in vitro with  -CD3 and -CD28 or alternatively with 5 µg/mL FV DbGagL peptide and -CD28 to detect intracellular IFN. Comparable results were obtained with both stimulation protocols. For the granzyme B and CD107a staining, harvested cells were analyzed directly ex vivo.
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